Description

pTWIN2 is an E. coli expression vector which can be used with the IMPACT™ Kit (NEB #E6901) (1). A polylinker in the vector is designed for the in-frame fusion of a target gene between the modified Ssp DnaB (2) and Mth RIR1 inteins (3). The presence of the chitin binding domain from Bacillus circulans (4,5) facilitates purification. pTWIN vectors are designed for protein purification or for the isolation of proteins with an N-terminal cysteine and/or a C-terminal thioester (1). The double-stranded vector is 7192 base pairs in length.

Advantages and Features

Features

A pBR322 derivative.

The SapI sites are recommended for directional cloning of both the 5´ and 3´ ends of an insert.

Expression of the fusion gene is under the control of the T7 promotor (8) and is regulated by IPTG due to the presence of a lacI gene.

Origin of DNA replication from the bacteriophage M13 allows for the production of single-stranded DNA by helper phage superinfection of cells bearing the plasmid.

Thiol-induced cleavage of the Mth RIR1 intein is dependent on the amino acids adjacent to the intein. The amino acid Gly at the C-terminus of the target protein is recommended for use with this intein.

Controllable cleavage of the Ssp DnaB intein is dependent on the amino acids adjacent to the intein. The amino acid sequence CRA or GRA at the N-terminus of the target protein is recommended for use with this intein.

Safety Data Sheet

Legal and Disclaimers

This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).

While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.

For more information about commercial rights, please contact NEB's Global Business Development team at gbd@neb.com.

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

Research Use Only

The buyer and user have a non-exclusive sub-license to use this system for any component thereof for RESEARCH PURPOSES ONLY, based upon agreement to the following assurances.

Transfer of this vector in host cells that contain the cloned copy of the T7 gene 1 to third parties is explicitly prohibited.

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